Method for affecting intracellular and extracellular electric and magnetic dipoles

ABSTRACT

A treatment of infectious diseases and enhancement of genetic engineering processes is described comprising the use of external electromagnetic energy to alter electric and magnetic dipoles intracellularly. 
     The process comprises introducing minute particles into the interior of cells in a host organism. These particles are capable of affecting the intracellular conductivity, dielectric properties, dipole content and membrane characteristics of the cell and nucleus. The particles are introduced intravenously, intra-arterially, and/or intra-lymphatically and the organism is then exposed to an alternating electromagnetic field to introduce energy into the cells.

FIELD OF THE INVENTION

This application is a continuation-in-part of U.S. patent applicationSer. No. 627,536 filed July 3, 1984 and now U.S. Pat. No. 4,767,611.

This invention relates to the field for affecting intracellular andextracellular electric and magnetic dipoles and more specifically to amethod for affecting intracellular and extracellular electric andmagnetic dipoles for use in the treatment of cancer and other diseases,including infectious diseases.

BACKGROUND OF THE INVENTION

The increased sensitivity of cancer cells as compared to normal cells toincreased temperature has been noted for some time. Cancer cells,because of their higher rate of metabolism, have higher restingtemperatures compared to normal cells. The normal resting temperature ofthe cancer cell is known to be 37.5° Centigrade, while that of thenormal cell is 37° Centigrade. Another physical characteristic thatdifferentiates the cancer cells from the normal cells is that cancercells die at lower temperatures than do normal cells. The temperature atwhich a normal cell will be killed and thereby irreversibly will beunable to perform normal cell functions is a temperature of 46.5°Centigrade) on the average. The cancer cell, in contrast, will be killedat the lower temperature of 45.5.° Centigrade. The temperature elevationincrement necessary to cause death in the cancer cell is determined tobe at least approximately 8.0° Centigrade, while the normal cell canwithstand a temperature increase of at least 9.5° Centigrade.

In attempts to solve the existing problem as to how to get energy intothe cancer cells without affecting the normal cells, efforts have beenmade to couple electromagnetic fields extracellularly to tissues toattempt to induce heating. However, these efforts have not beeneffective largely because of the inability to differentiate the cancercells form the normal cells.

The electromagnetic field interacts with tissue in several ways Thereare displacement currents due to the drift of electrons, polarization ofatoms or molecules to produce dipoles and the interaction with dipolesalready present. The coupling of electromagnetic energy to the tissuedepends on the electrical conductivity (σ) and the dielectric constant(ε). The power imparted to the tissue depends on the square of theamplitude of the field and the coupling constant to the tissue. Thedielectric properties of the material depend on its composition andstructure (i.e ions, polar molecules, etc.).

In general:

    ε=ε'-jε"

where

ε'=real component related to energy stored in the material in electricfields

ε.increment.=imaginary component related to loss in the form of heat=##EQU1##

σ=conductivity so that conductivity is related to the amount of heatloss Often as there is an increase in frequency, ε' decreases due toless ordering and ε" increases.

In tissue a plot of the dielectric constant as a function of frequencyoften shows three dispersions. Each dispersion is related to a specificphenomenon. The α dispersion (at≈80-100 Hz) is due to the interaction ofthe charges on the cell surface with the ions in solution and theimpedence if the membrane system.

The β dispersion (at≈50 KHz) is related to the cell membrane'sinsulation of the H₂ O. Above 10 GHz the γ dispersion is due to the H₂ Oand electrolyte solution. To overcome these problems the common practiceis to use a frequency>>1 KHz to short out the membrane effects and todeliver energy to the cytoplasm.

Consequently, frequencies greater than 1 KHz and usually greater than 1MHz are utilized to overcome problems with the cell membrane and deliverenergy to the cell. Traditionally, frequencies of 13 MHz or 2450 MHz areused. However, the problem remains that at these high frequencies notonly are the cancer cells affected but the normal cells are alsoaffected and consequently one is limited in the amount of energy whichcan be delivered to the cancer cells.

The present invention seeks to overcome this problem, by modifying theintracellular environment to allow the use of lower frequencies ifpossible and to enhance the effect in the cancer cells without affectingthe normal cells.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A: Illustrates the measurements of impedence in tissue as afunction of frequency for Tumor tissue (control and treated) and Livertissue (control and treated).

FIG. 1B: Demonstrates the A-C conductivity Tumor/Liver ratio (controland particle living tissue) as a function of frequency.

FIG. 2A: Illustrates the A-C conductivity ratio Tumor/Liver (control andparticle containing tissue) in a second series of animals as a functionof frequency.

FIG. 2B: Illustrates the difference in impedence between the Tumor andthe Liver in the treated animals as compared to the Control animals as afunction of frequency.

OBJECT OF THE INVENTION

The present invention comprises the introduction of minute particlesinto cells of a host organism to alter the intracellular environment andto enhance the effect of external electromagnetic energy on the cellsconsequently affecting the cells without affecting the normal cells.

DESCRIPTION OF THE INVENTION

The use of particles to alter the energy level in the cell by theabsorption of electromagnetic energy by the particle has been disclosedby the applicant in his U.S. Pat. Nos. 4,106,488; 4,136,683; 4,303,636;and 4,359,453. The present invention seeks to use particles to affectthe absorption of energy by the cell itself in response to an externalalternating electromagnetic field. The implementation of this presenttreatment method in part utilizes inventive aspects which are thesubject of other applications for U.S. Letters Patent by the sameinventor as recited- hereinafter. For example, a fuller understanding ofthe technology underlying the Gordon treatment reveals the operation ofsubtle mechanisms which can themselves become a contributing factor inthe course of treatment and are incorporated herein by reference. Theselection of particle compositions for use in this present invention asdisclosed in the applicant's above described U.S. Patents and asdisclosed in copending and commonly assigned applications Ser. Nos.418,298; 464,870 including C.I.P; 522,941 including C.I.P. 535,390;524,844; and 561,811 of the same inventor, are incorporated herein byreference.

Below several MHz the transmission of energy directly to the cell by anexternal alternating electromagnetic field is affected by thecharacteristics of the cell membrane. The charge accumulation on themembrane from intracellular and extracellular fluids accounts for thedielectric polarization of the membrane. The intracellular andextracellular electrolyte solution accounts for the conductance.

In the prior Robert T. Gordon U.S. Pat. Nos. 4,303,636; 4,106,488 and4,359,455, a high frequency magnetic field is employed to have a directeffect on the particles so that diseased tissue can be killed by thermaleffects due to hysteresis loss form the particles themselves when thefield is relaxed In contrast to this phenomena, the particles of thepresent invention are employed to alter the environment such as themagnetic dipoles charge accumulation and conductivity both intra andextra cellularly of a host organism.

The altered environment in turn produces thermal effects (an increase intemperature) when subjected to an alternating magnetic field atrelatively low frequencies. These thermal effects are not due to ahysteresis loss from the particles. The frequency range employedaccording to the present invention which is lower than that of theaforesaid Gordon patents thereby reduces the power requirements. Thedesign and timing of treatment will differ because of the lowerfrequency ranges employed in the treatment methods of the presentinvention. One of the additional advantages of the present inventionresides in the fact that coil apparatus does not have to be used as wasthe case in the Gordon patents noted above, the instant inventioncapable of being practiced inter alia with capacitor plates fordeveloping the frequencies required for treatment.

In the Gordon patents noted above, optimum results were obtained by themetabolism of the particles by the cells. In the present inventioneffective treatment may be obtained outside the cell environment sincethe membrane surface dipoles are affected by the particles. The presentinvention, however, also applies to affecting the dielectric,conductivity and frequency dependent dispersion curves both extracellularly as well as intra cellularly (i.e., those instances where theparticles are metabolized or absorbed by the cells of a host organismwhether the cells are diseased cells or normal cells).

Through the introduction of particles in the cancer cells theintracellular conductivity can be altered as well as the chargeaccumulation on the cell membrane This enhancement of conductivity aswell as the alteration in membrane events allows two benefits Thealteration in cell membrane characteristics enables the delivery ofenergy intracellularly at a lower frequency due to the effect on thecharge accumulation on the cell membrane. The increased conductivity ofthe cell allows more energy to be delivered and a better coupling at thegiven frequency.

In addition, the effect on the normal cells is greatly reduced becausethe energy requirements are less, the normal cell membrane still acts asa barrier at this frequency and the frequency to be utilized can belowered.

Consequently, through the alteration of the intracellular environmentand the membrane effects at a given frequency, more energy can bedelivered to the cancer cells, a lower frequency can be utilized sincethe cancer cell membrane will start to short out at a lower frequencyand the effect on the normal cells is reduced due to its intact cellmembrane characteristics.

EXAMPLES

In this original study 12 Sprague-Dawley rats with spontaneous mammarytumors were utilized. Six of the animals were injected with 2 cc. ofFeTPPS₄ Acetate in a dosage of 10 mg/cc. The animals were observed forany side effects and at 48 hours biopsies were performed of the tumor,liver, spleen, kidney, heart, and lung. These biopsies were performed onboth the control and injected animals.

Measurements of conductivity were then performed at 37° C. in the livingtissue of the dielectric properties by means of a Bonton Impedencemeasuring device and the status of the electric and magnetic dipoles.The results are reported in FIGS. 1A, 1B, 2A and 2B which illustrate thedifference in dielectric properties between the treated and untreatedtumor as compared to a normal organ such as the liver;

FIGS. 1A and 2B show the change in impedence with frequency for severaltissues. The particle containing tumor tissue appears more conductivethan the control tumor tissue or the liver tissue.

In the tumor containing the particles the conductivity increases as doesthe magnetic susceptibility (Gordon et al. in press). However, in theliver tissue due to differences in metabolism and processes of theparticle, the conductivity decreases and the magnetic susceptibilitydecreases (Gordon et al. in press). This difference in processing of theparticle system is designed to maximize the ability to differentiallyaffect the tumor cells with minimal or no effect to the normal cells.Differences in processing of systems by different tissues is seen by thedifference of fluorescence in the same amount of porphyrins in differenttissues.

FIGS. 1B and 2A illustrates another aspect of the present invention.FIGS. 1B, and 2A show the plot of the conductivity ratio of Tumor/Liverof particle containing tissue versus control tissue. These studies wereperformed with living tissue having been injected intravenously withFeTPPS₄ -acetate. The data suggests for this particle system a maximumat 500 Hz. However, at 10 Hz and also at 200 KHz-500 KHz the T/Lconductivity for the particle containing tissue is 2× that of the normaltissue which can prove useful.

In addition, the ratio for particle containing tissue is higher at lowerfrequencies as suggested by the shift of the left hand hump of theparticle containing curve to the left. This data supports the use oflower frequencies in the particle containing systems.

The dielectric properties and conductivity of materials are temperaturedependent. The membrane charging time constant and frequency ofdispersion often vary 2% per °C. The frequency dependent dispersioncurves of materials vary with temperature. A further innovation of theinstant invention is to utilize the dielectric properties, conductivity,and frequency dispersion curves of the living tissue to identify thetemperature of that tissue and follow the treatment process.

The dielectric properties change with the metabolism rate in the celland consequently measurement of these properties allows knowledge notonly of the temperature but also the state of metabolism in the cell andallows a means of following changes in the state of metabolism.

Similar results are available from the other tissues studied and readilydemonstrate the effect of the particles on the dielectric propertiesconductivity and electric dipoles of certain tissues and thedifferential between the tumor tissue and normal tissue.

As disclosed by the applicant in his U.S. Pat. No. 4,136,683 and asdisclosed in copending and commonly assigned application Ser. No.535,390 (C.I.P. to application Ser. No. 522,941) of the same inventorand incorporated herein by reference, this present invention can be usedto create a three-dimensional temperature map of the body. In addition,the measurement of these properties allows one to follow thedistribution of the particles in the body by following the change in thedielectric properties, conductivity, and frequency dispersion curvesboth before and after ingestion of the particles.

Molecules in a cell can be affected if μE≧kT (where μ is dipole moment,E is the field strength, k is the Boltzman constant, and T is absolutetemperature). Consequently by introducing the particles and increasingthe relative dipole moment in the cell the direct effects on moleculesin the cell can be enhanced even beyond thermal effects. Therefore, thispresent invention may directly affect the molecules in the cell.

Through these processes the dielectric properties across the membranecan be affected including the stimulation and/or alteration of nerveimpulses and/or electrical events.

The ionic environment around the surface of the particle by becomingpolarized can produce increased dielectric properties as well. Inaddition, membrane effects with the anionic proteinaceious materialwhich accumulates around the cell can produce local effects.

When you have a mixture with different dielectric properties relaxationphenomenon will occur not at a single frequency, but over a wide rangeof frequencies. The curve is broadened due to interactions in themixture. Inclusion of material of low dielectric constant will lower thedielectric constant of the mixture. Therefore, the addition of particlesto the inside of the cell broadens the frequency response ofintracellular structures as compared to the other cells and structures.Particle geomentry also affects the frequency response. Consequently,the presence of the particles allows for a differential affect onsubcellular structures.

The choice of particle type, size and shape can be highly significant toeffective treatment, particularly where subcellular localization orother subtle differentiations in metabolic activity, for example, areconveniently utilized to maximize particle uptake and absorption.Suitable particles and exemplifications of selection parameters aredisclosed and examined in copending and commonly assigned applicationSer. No. 535,390 of the same inventor, incorporated herein by reference.

The particles may be chosen from ferromagnetic, paramagnetic, ordiamagnetic particles. The particle systems include metalloporphyrins,Fe₂ O₃, metal-metalloporphyrins and particularly useful particlesincluding both inorganic elements and compounds as well as metalcontaining organic compounds. Inorganic elements and compoundsparticularly well suited, owing to their favorable magnetic parameters,comprise elements such as dysprosium, erbium, europium, gadolinium,holmium, samarium, terbium, thulium, ytterbium or yttrium and compoundsthereof such as dysprosium sulfate, erbium sulfate, europium oxide,europium sulfate, gadolinium oxide, gadolinium sulfate, holmium oxide,samarium sulfate, terbium oxide, terbium sulfate, thulium oxide,ytterbium sulfide, yttrium oxide, yttrium sulfate, yttrium ferrioxide(Y₃ Fe₅ O₁₂), yttrium aluminum oxide (Y₃ Al₅ O₁₂), other dimetalliccompounds such as dysprosium-nickel, dysprosium-cobalt, gadolinium-iron,ytterbium-iron, cobalt-samarium, gadolinium-yttrium, anddysprosium-gallium, and actinide series elements and compounds thereof.

Metal containing-organic molecules useful for the application describedabove, comprise particles of iron-dextrans such as FeOOH-dextrancomplexes and other dextran metal complexes wherein the metal isselected from the group comprising cobalt, iron, zinc, chromium, nickel,gallium, platinum, manganese and rare earth metals such as dysprosium,erbium, europium, gadolinium, holmium, samarium, terbium, thulium,ytterbium and yttrium, other dimetallic compounds such asdysprosium-nickel, dysprosium-cobalt, gadolinium-iron, ytterbium-iron,cobalt-samarium, gadolinium-yttrium, and dysprosium-gallium and ironsuch as Fe₂ O₃ particles, Fe₃ O₄ particles and FeOOH particles and Fe₂O₃ -dextran complexes, Fe₃ O₄ -dextran complexes, and FeOOH dextrancomplexes, and actinide series elements and. compounds, ferric ammoniumcitrate, and various iron transporting and chelating compounds such asenterochelin, transferrin, metallothionein, hydroxamates, phenolates,ferrichromes, desferri-ferrichromes, ferritin, ferric mycobactins andiron-sulfur proteins such as ferredoxin and rubredoxin and transferrinas well as transferrin compounds and complexes.

Particularly appropriate metal-containing organic structures for usewith the present invention are the porphyrins such as etioporphyrins,mesoporphyrins, uroporphyrins, coproprophyrins, protoporphyrins, anddicarboxylic acid containing prophyrins and substituted prophyrins suchas tetraphenylporphyrin sulfonate (TPPS). Especially advantageousprotoporphyrins comprise hematoporphyrins, chlorophylls, andcytochromes. In addition to the naturally occurring protoporphyrinswhich possess either iron or magnesium containing moieties, mixed-metalor di-metal hybrid prophyrins may also be prepared. For example, bysubstituting an alternative metal for the iron in hematoporphyrin, theadvantages of the porphyrin moiety (e.g., in terms of specificity oflocalization is retained while the unique magnetic properties of the newmetal enhance the sensitivity of the substituted molecule. Suitablemetals for purposes of substitution comprise cobalt, iron, manganese,zinc, chromium, gallium, nickel, platinum and rare earth series ofmetals such as dysprosium, erbium, europium, gadolinium, holmium,samarium, terbium, thulium, ytterbium and ytterium, dimetallic compoundssuch as dysprosium-nickel, dysprosium-cobalt, gadolinium-iron,ytterbium-iron, cobalt-samarium, gadolinium-yttrium, dysprosium-galliumand actinide series elements and compounds thereof. The substitutedporphyrins are then optionally reacted with dextran to form ametal-containing porphyrin dextran complex in particle form. Suitableporphyrin acceptors comprise any dicarboxylic acid containing prophyrinsuch as protoporphyrins (e.g., hematoporphyrins) and the like.

The substitution reaction is carried out in vitro by reacting thedesired metal with the desired porphyrin in the presence of the enzymeferrochelatase (E.C. 4.11.1.1). Reaction conditions as described byJones and Jones (Biochem. J. 113:507-14, 1969) or Honeybourne, et al.(FEBS Lett.: 98:207-10, 1979) are suitable.

Additional particle systems particularly suited to use in this presentinvention include Fe₃ O₄ -transferrin dextran, metal-transferrin(transition, rare-earth), metalloporphyrin-transferrin,antibody-ferritin-particles, antibody-ferritin-transferrin particles,antibody-transferrin particles, metal -porphyrin-metal complexes,metallothionein particles, and lectin particles. Useful particle systemsfor use in this present invention further comprise: Where particle=Fe₃O₄, transition metal, rare-earth metal, metalloporphyrin, etc. as wellas ferromagnetic and paramagnetic particles.

One magnetic characteristic known to be temperature dependent ismagnetic susceptibility. Magnetic susceptibility is measured by theratio of the intensity of magnetization produced in a substance to themagnetizing force or intensity of the field to which it is subjected.This magnetic characteristic is routinely measured by magnetometerdevices such as a vibrating magnetometer or a flux gate magnetometer.Therefore, by measuring the magnetic susceptibility of particles atvarious temperatures, it is quite simple to calibrate the magnetometerequipment so that when it measures the magnetic susceptibility of theparticles a simple calibration will indicate the exact correspondingtemperature of the particle.

By way of illustrating the increased magnetic susceptibility of some ofthe elements or compounds described above, the following table isprovided:

    ______________________________________                                        Element or Compound                                                                            Temp (K)  Mag.Sus.(10.sup.6 cgs)                             ______________________________________                                        Iron Oxide (ref.)                                                                              293        +7,200                                            Dysprosium Oxide 287.2     +89,600                                            Dysprosium Sulfate                                                                             291.2     +92,760                                            Octahydrate                                                                   Erbium Oxide     286       +73,920                                            Erbium Sulfate Octahydrate                                                                     293       +74,600                                            Europium         293       +34,000                                            Europium Oxide   298       +10,100                                            Europium Sulfate 293       +25,730                                            Holmium Oxide    293       +88,100                                            Holmium Sulfate Octahydrate                                                                    293       +91,600                                            Terbium          273       +146,000                                           Terbium Oxide    288.1     +78,340                                            Terbium Sulfate                                                               Octahydrate      293       +76,500                                            Thulium          291       +25,500                                            Thulium          296.5     +51,444                                            Ytterbium Sulfide                                                                              292       +18,300                                            ______________________________________                                    

Thus, the enhanced magnetic characteristics displayed by the particlesof the subject invention results in an increase in an electromagneticfield thereby increasing the overall sensitivity and control of themodalities for the improvement of the instant invention techniques andfor the resultant effects.

Magnetic susceptibility has also been used heretofore in connection withthe treatment protocol as disclosed in U.S. Pat. No. 4,163,683 of thesame inventor, where magnetic susceptibility measurements are correlatedwith temperature (an interdependent variable) in accomplishing therelated induction heating step controllably. There is no recognition,however, that the values for magnetic susceptibility, independent of theinduction heating step or the imposition of an electromagnetic field canbe usefully correlated (to maximization of particle concentration withtime to optimize treatment effectiveness, as demonstrated herein.

A further benefit is derived from the fact that some particlecompositions comprise a ferromagnetic, paramagnetic, or diamagneticcomponent integrated into a cell or organelle specific molecularstructure, thereby permitting efficient targeting and delivery of saidparticles to specific intracellular compartments such as mitochrondria,chloroplasts, nuclei, vacuoles, and the like.

This present invention is also applicable to be used in the treatment ofatherosclerotic lesions. Descriptions of various aspects of treatmentfor atherosclerotic lesions is disclosed in U.S. Pat. No. 4,359,453 ofthe same inventor, incorporated herein by reference.

Further application of this present invention includes the treatment ofdisease processes, including infectious diseases and organismscomprising Salmonella, Kelbsiclla, Escherichia, Clostridium,Mycobacterium, Pseudomonas, Peptostreptococcus, Phycomyces, Candida,Ustilago, Entamocba, Trypanosoma, Leishmania and RNA viruses.Descriptions of various aspects of treatment for the above described isdisclosed in copending and commonly assigned application Ser. Nos.464,870; 468,644; and 524,844 of the same inventor, incorporated hereinby reference.

Thus the invention deals with the use of intracellular particles toenhance the ability to treat infectious diseases. These effects includechanges in the cell membrane, receptor sites and interaction withcellular components i.e. mitochondria. It is clear that an effect on thecell membrane and its interrelationship with the mitochondria and thestate of energy metabolism in the cell will affect the ability of thecell to resist an infectious agent.

In regard to the aforesaid treatment of infectious diseases, the changesin cell membrane receptors sites and interaction with cellularcomponents, i.e. mitochondria, are also clear. Alterations in any ofthese processes will affect the eneregy level and metabolic rate in thecell and will easily affect the ability of the cell to resist infectiousagents. As an example, the membranes of many bacteria releasesiderophores which gather iron essential to the bacteria's metabolism.Alteration of membrane events affects the ability of the bacteria togather the iron and hence affects their metabolism. Similarly,trypanosomes alter the transferrin receptors on cells in number andquality and alters the cell's ability to change energy levels and toresist infection. Therefore by interacting with the cell membranereceptors, one can control the ability of the trypanosomes to survive.Hence, membrane events can control the outcome of infectious diseaseprocess.

In addition, particle systems which are kept outside the cell may beutilized to alter membrane events and affect the frequency of responseand the energy transmission of the cancer cells and the normal cells Incertain circumstances these particles may be utilized to stabilize themembrane of normal cells and decrease their response to a field at agiven frequency.

A steady magnetic or electric field may be used to enhance the uptake ofparticles by the cells as well as enhancing the membrane and cytoplasmicalterations which occur and are fully disclosed and described incopending and commonly assigned application Ser. No. 535,390 of the sameinventor incorporated herein by reference. For Example, the applicationof the localized static mangetic or electric field may occurconcurrently with the application of an alternating, oscillating orpulsed electromagnetic field. That is to say, the localized staticmagnetic or electric field may be superimposed on the subject ofinterest while the alternating, oscillating or pulsed field is alsobeing applied.

Genetic Engineering processes and DNA-RNA modifications may also beaffected and enhanced by the processes described in this presentinvention. Descriptions of aspects of treatment for the above describedin disclosed in copending and commonly assigned application Ser. No.418,298 of the same inventor incorporated herein by reference.

These modification include changes in conformation, structure,alteration in binding sites and changes in histone binding. All of thesemodifications will affect the genetic engineering process. As anexample; when a portion of nucleic acid is introduced into a strand ofDNA, the conformation and binding sites available on the DNA greatlyinfluence whether the nucleic acid is able to join the DNA. The exactmodification depends on the desired result.

Further with regard to DNA, RNA modifications, the fact that changes inconformation, structure, alteration in binding sites and changes inhistone binding will affect the function of DNA-RNA interactions iswell-known to those "skilled in the art." The binding of DNA or nucleicacids to histones and proteins greatly affects the ability of the DNA tobe functionally active as well as to affect its conformation.

Ultrasound mechanisms may also be affected by the processes described inthis present invention to enhance the differentiation techniques. Thisis evidenced by the attenuation of ultrasound energy in particle systemssimiliar to iron porphyrin FeTPPS₄ i.e. (Hemoglobin). The orientation ofthe tissue is also important and can be utilized to maximize the effectof the E field or the H field.

Temperature measurements are taken in living tissue of the host organismand correlating the temperature readings to the low frequency magneticfield causing alteration in dielectric properties and/or conductivityand/or frequency dependent dispersion curves. Once the temperature iscorrelated with these measurements, (dielectric properties, conductivityand frequency dependent dispersion curves) these measurements are thenmade along three axes at right angles to one another in the hostorganism from which a three dimensional temperature map of the body isproduced by restructuring them in a three dimensional temperature modelby computer processes well known in the art.

The frequency of the magnetic field is selected to enhance thedielectric properties, conductivity and electric dipoles of the cells ofthe host organism and will vary depending upon the particles employed inthe host. The frequency, however, is adjusted so that thermal effectsthereby obtained are not due to hysteresis loss from the particlesthemselves but rather the alteration in conductibility, dielectricproperties and electric dipoles of the cells that are brought about bythe use of the particles of the present invention. Generally, the rangeof frequencies that may be employed will be anywhere from about 1 hz toabout 500 MHz; 1 Hz to about 100 MHz; 1 Hz to less than 13 MHz; 1 Hz toabout 100 KHZ; 1 hertz up to less than 50 kilohertz and especially fromabout 10 hertz up to about less than 50 kilohertz as well as anyfrequency within these ranges or range of frequencies within theaforesaid ranges.

The present invention, therefore, will be practiced at the abovefrequencies and the copending applications incorporated herein byreference will give the person of ordinary skill in the art a disclosureof how to practice the present invention with the exception that thefrequencies described above will be employed in lieu of those utilizedin such copending applications.

To further illustrate the operation of the subject procedure, thefollowing treatment scenario is provided.

Reference herein to tissue, organ or cell population is intended in itsmost embracive and comprehensive sense, referring in general to theregion of the host organism affected by the invasive abnormality, or thetreatment region, as the context requires.

The subject receives an intravenous injection of a colloidally suspendedparticle such as iron porphyrin (FeTPPS₄) at a dosage of 2-10 mg/kg.After 48 hours, the subject is exposed to an alternating electromagneticfield at a frequency of 1 Hz to 100 MHz in this case 500 Hz for a periodof approximately 10-20 minutes. The alternating electromagnetic fieldmay be applied via a coil arrangement or via capacitor plates or viaelectrodes in the tissue or any suitable means available in the state ofthe art, and consistent in application to this present invention. Theprocess may be repeated as is necessary.

In summary, the introduction and absorption of minute particles into thecell alters the intracellular environment and the charge accumulation onthe membrane. Consequently, lower power levels may be used to transmitenergy into the cell, lower frequencies may be used because of thealteration in membrane events and the effect on normal cells is greatlyreduced because of the above as well as the state of the normal cell'smembrane. In addition, modification can be performed by using particlesto alter the extracellular environment as well.

In addition, since radiofrequency fields can affect particles by causingreversible or irreversible changes in the particles, i.e.magnetostrictive induced vibrations, by affecting the particles with analternating electromagnetic field in the range 1 Hz to 500 MHz eitherprior or during treatment, the particles can be made more or lessresponsive to the field. This alternating field can produce acousticchanges in the particle and affect the cell and subcellular structures.

What is claimed is:
 1. A process for the treatment of diseased cells inat least one region in the tissue of a host organism containing saiddiseased cells and living normal cells without substantially damagingsaid living normal cells comprising:providing to said host organismminute particles less than about 1 micron capable of being taken up bysaid diseased cells; selecting said particles to affect intracellularand extracellular events for the enhancement of treating infectiousdiseases produced by organisms selected from the group consisting ofSalmonella, Klebsiella, Escherichia, Clostridium Mycobacterium,Pseudomonas, Peptostreptococcus, Phycomyces, Candida, Ustilago,Entamoeba, Trypanosoma, Leishmania and RNA viruses by the selection ofsaid particles; allowing said particles to effect at least one eventcomprising intracellular events and membrane events in said tissue;subjecting said organism to a relatively low frequency alternating,oscillating, or pulsed electromagnetic field to provide energy to saiddiseased cells and selectively heat said diseased cells wherein saidparticles are selected from the group consisting of: a) cobalt, zinc,iron, chromium, nickel, platinum, rare earth metals and compoundsselected from the group consisting of dysprosium, erbium, europium,gadolinium, holmium, samarium, terbium, thulium, ytterbium, yttrium;dysprosium sulfate, erbium sulfate, europium oxide, europium sulfate,gadolinium oxide, gadolinium sulfate, holmium oxide, samarium sulfate,terbium sulfate, thulium oxide, ytterbium sulfide, yttrium oxide,yttrium sulfate, yttrium ferrioxide (Y₃ Fe₅ O₁₂), yttrium oxide (Y₃ Al₅O₁₂), dysprosium-nickel, dysprosium-cobalt, gadolinium-iron,ytterbium-iron, cobalt-samarium, gadolinium-ytterbium,dysprosium-gallium, and actinide series elements and compounds thereof;b) dextran metal complexes wherein said metal is selected from the groupconsisting of cobalt, zinc, chromium, iron, gallium, manganese, nickel,platinum, dysprosium, erbium, europium, gadolinium, holmium, samarium,terbium, thulium, ytterbium, yttrium, dysprosium-nickel,dysprosium-cobalt, gadolinium-iron, ytterbium-iron, cobalt-samarium,gadolinium-yttrium, and dysprosium-gallium; c) iron transporting andchelating compounds selected from the group consisting of ferricammonium citrate, enterochelin, transferrin, metallothionein,hydroxamates, phenolates, ferrichromes, desferriferrichromes, ferritin,ferric mycobactins, ferredoxin and rubredoxin; d) porphyrins selectedfrom the group consisting of etioporphyrins, meso-porphyrins,uroporphyrins, coproporphyrins, protoporphyrins, dicarboxylic acidcontaining porphyrins, tetraphenylporphyrin sulfonate, hematoporphyrins,chlorophylls, and cytochromes; e) and combinations of the materials ofsaid sub-paragraphs a), b), c) and d).
 2. A process comprising selectingparticles of less than about 1 micron to affect intracellular andextracellular events for the enhancement of Genetic Engineeringprocesses or for DNA-RNA modifications by the selection of saidparticles comprising, introducing said particles into a host organismand subjecting said host organism to a relatively low frequencyalternating, oscilating or pulsed electromagnetic field to provideenergy to said organism and enhance said Genetic Engineering processwherein said particles are selected from the group consisting of:a)cobalt, zinc, iron, chromium, nickel, platinum, rare earth metals andcompounds selected from the group consisting of dysprosium, erbium,europium, gadolinium, holmium, samarium, terbium, thulium, ytterbium,yttrium; dysprosium sulfate, erbium sulfate, europium oxide, europiumsulfate, gadolinium oxide, gadolinium sulfate, holmium oxide, samariumsulfate, terbium sulfate, thulium oxide, ytterbium sulfide, yttriumoxide, yttrium sulfate, yttrium ferrioxide (Y₃ Fe₅ O₁₂), yttrium oxide(Y₃ Al₅ O₁₂), dysprosium-nickel, dysprosium-cobalt, gadolinium-iron,ytterbium-iron, cobalt-samarium, gadolinium-ytterbium,dysprosium-gallium, and actinide series elements and compounds thereof;b) dextran metal complexes wherein said metal is selected from the groupconsisting of cobalt, zinc, chromium, iron, gallium, manganese, nickel,platinum, dysprosium, erbium, europium, gadolinium, holmium, samarium,terbium, thulium, ytterbium, yttrium, dysprosium-nickel,dysprosium-cobalt, gadolinium-iron, ytterbium-iron, cobalt-samarium;gadolinium-yttrium, and dysprosium-gallium, Fe₃ O₄ Fe₂ O₃, Fe₂ O₃,FeOOH: c) iron transporting and chelating compounds selected from thegroup consisting of ferric ammonium citrate, enterochelin, transferrin,metallothionein, hydroxamates, phenolates, ferrichromes,desferriferrichromes, ferritin, ferric mycobactins, ferredoxin andrubredoxin; d) porphyrins selected from the group consisting ofetioporphyrins, meso-porphyrins, uroporphyrins, coproporphyrins,protoporphyrins, dicarboxylic acid containing porphyrins,tetraphenylporphyrin sulfonate, hematoporphyrins, chlorophylls, andcytochromes; e) and combinations of the materials of said sub-paragraphsa), b), c) and d).
 3. A process for the treatment of diseased cells inat least one region in the tissue of a host organism containing saiddiseased cells and living normal cells without substantially damagingsaid living normal cells comprising:providing to said host organismminute particles less than about 1 micron capable of being taken up bysaid diseased cells; selecting said particles to affect intracellularand extracellular events for the enhancement of treating infectiousdiseases produced by organisms selected from the group consisting ofSalmonella, Klebsiella, Escherichia, Clostrididium, Mycobacterium,Pseudomonas, Peptostreptococcus, Phycomyces, Candida, Ustilago,Entamoeba, Trypanosoma, Leishmania and RNA viruses by the selection ofsaid particles; allowing said particles to effect at least one eventcomprising intracellular events and membrane events in said tissue;subjecting said organism to a relatively low frequency alternating,oscillating, or pulsed electromagnetic field to provide energy to saiddiseased cells and selectively heat said diseased cells wherein saidparticles are selected from the group consisting of: a) ferromagnetic,paramagnetic and diamagnetic elements, inorganic compounds, organiccompounds and combinations thereof,metalloporphyrins, Fe₂ O₃, FeOOH, andmetal metalloporphyrins.
 4. A process comprising selecting particles ofless than about 1 micron to affect intracellular and extracellularevents for the enhancement of Genetic Engineering processes or forDNA-RNA modifications by the selection of said particles comprising,introducing said particles into a host organism and subjecting said hostorganism to a relatively low frequency alternating, oscilating or pulsedelectromagnetic field to provide energy to said organism and enhancesaid Genetic Engineering process wherein said particles are selectedfrom the group consisting of:a) paramagnetic and diamagnetic elements,inorganic compounds, organic compounds and combinations thereof,metalloporphyrins, Fe₂ O₃, FeOOH, and metal metalloporphyrins.
 5. Theprocess of claims 1, 2, 3 or 4 wherein said alternating, oscillatingand/or pulsed electromagnetic field is between about 1 Hz to about 100MHz.
 6. The process of claims 1, 2, 3, or 4 wherein said alternating,oscillating and/or pulsed electromagnetic field is produced by apparatuscomprising a coil, capacitor plates or electrodes in the said hostorganism.
 7. The process of claims 1, 2, 3 or 4 wherein said particlesare selected to affect the intracellular conductivity, dielectricproperties, charge accumulation on the membrane, membrane conductance,membrane capacitance and the electric dipole environment of cells. 8.The process of claims 1, 2, 3 or 4 wherein the natural occurring metalmoiety of said porphyrin is optionally substituted with a metal selectedfrom the group comprising cobalt, zinc, chromium, gallium, iron,manganese, nickel, platinum, dyprosium, erbium, europium, gadolinium,holmium, samarium, terbium, thulium, ytterbium, yttrium,dyprosium-nickel, dyprosium-cobalt, gadolinium-iron, yttrebium-iron,cobalt-samarium, gadolinium-yttrium, and dyprosium-gallium;andcombinations thereof.
 9. The process according to claims 1, 2, 3 or 4wherein said iron transporting, iron chelating and porphyrin compoundsare chemically complexed with dextran.
 10. The process according toclaims 1, 2, 3 or 4 wherein said compound is chemically complexed withan antibody.
 11. The process of claims 1, 2, 3 or 4 for heating saidhost organism comprising decreasing the power level at a given frequencyof said field by altering intracellular environment and membranecharacteristics in said tissue by the provision of said particles tosaid host.
 12. The process of claims 1, 2, 3 or 4 for heating saiddiseased cells comprising decreasing the frequency of said field byaltering the intracellular environment and membrane characteristics insaid tissue by the provision of said particles to said host.
 13. Theprocess of claims 1, 2, 3 or 4 comprising applying a localized staticmagnetic field to said host organism to aid in the intracellular uptakeand energy absorption of the electric or magnetic dipoles eitherintroduced or already present in said host organism.
 14. The process ofclaims 1, 2, 3 or 4 wherein a localized static magnetic or electricfiled is applied to said host organism after providing said particles tosaid host organisms but prior to or during the application of saidalternating, oscillating and/or pulsed electromagnetic field to enhancethe intracellular of energy and the energy-absorption responsiveness ofsaid particles.
 15. The process of claim 13 wherein a localized staticmagnetic or electric field is applied to said host organism prior toand/or during the application of said alternating, oscillating and/orpulsed electromagnetic field to enhance the intracellular energy uptakeand energy absorption of the electric or magnetic dipoles eitherintroduced or already present in said tissue.
 16. The process of claim13 wherein the static magnetic or electrical field is between 100 gaussand 80 kilogauss.
 17. The process of claims 1 or 3 wherein saidparticles are introduced into the extracellular environment of saidtissue to alter membrane events and potentiate energy delivery to saiddiseased cells and/or reduce energy delivery to said normal cells. 18.The process of claims 1 or 3 wherein particles are introducedintravenously, intra-arterially, intra-lymphatically, and or locally.19. The process of claims 1, 2, 3 or 4 wherein an alternatingelectromagnetic filed between 1 Hz and 500 MHz is used to affect saidparticles and make them more or less responsive to an excitingalternating electromagnetic field produced by magnetostrictive inducedvibrations applied to said host organism.